Applications

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application

Abreviews

Notes

ICC/IF

Use a concentration of 0.65 µg/ml. PubMed: 19001505

Flow Cyt

Use a concentration of 0.65 µg/ml. PubMed: 19001505Ab81216-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ChIP

Use at an assay dependent concentration. PubMed: 18332116

WB

Use a concentration of 0.2 - 2 µg/ml. Predicted molecular weight: 37 kDa. Detects Bmi-1 in RIPA lysates from U2OS cells. U2OS cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with ab14389 0.2ug/ml. Proteins were visualized using a goat anti-mouse IgG labeled with HRP and a chemiluminescence detection system.

ICC

Use at an assay dependent concentration.

IP

Use at an assay dependent concentration.

IHC-Fr

Use at an assay dependent concentration. PubMed: 17210912

IHC-P

1/100.

Target

FunctionComponent of the Polycomb group (PcG) multiprotein PRC1 complex, a complex required to maintain the transcriptionally repressive state of many genes, including Hox genes, throughout development. PcG PRC1 complex acts via chromatin remodeling and modification of histones; it mediates monoubiquitination of histone H2A 'Lys-119', rendering chromatin heritably changed in its expressibility. In the PRC1 complex, it is required to stimulate the E3 ubiquitin-protein ligase activity of RNF2/RING2.

Sequence similaritiesContains 1 RING-type zinc finger.

Post-translationalmodificationsMonoubiquitinated (By similarity). May be polyubiquitinated; which does not lead to proteasomal degradation.

Anti-Bmi1 antibody [1.T.21] - ChIP Grade images

SK-N-SH cells were fixed in 4% paraformaldehyde, permeabilized in 0.55 Triton-X100 and incubated for 1 hour with ab14389 (1/200). The Bmi1 staining is shown in red. The cells were counterstained with DAPI (blue). 100x magnification.

ab14389 staining Bmi1 in human head and neck squamous cell cancers (HNSCC) by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).Sections were deparaffinized in xylene, dehydrated through graded alcohols, and placed in 0.1% hydrogen peroxide to quench any endogenous peroxidase activity. A 5 minute, 750 W microwave pretreatment in citrate buffer (pH 6.0) was repeated 4 times and followed by treatment with 10% normal rabbit serum for 30 minutes to block nonspecific antibody binding. The slides were then incubated with ab14389 at a 1/100. Primary antibody incubation was followed by three washes with PBST and incubation with fluorescently-labeled Cy2 (1/250) secondary antibody for 2 hours. Nuclei were counterstained with DAPI. Finally, sections were rinsed with PBST, dehydrated through sequential washes in 50%, 70%, 95%, and 100% ethanol and then cleared in xylene. Slides were mounted with DPX mounting media and allowed to dry overnight.